Using a method of retroviral gene transduction we have been able to generate conditionally immortalized long-term hematopoietic stem cell lines from primary mouse blood stem cells. Phase I of this project included applying this technology to generate human cell lines. In addition, the project included testing a non-viral method to generate the stem cell lines from mice. Phase II of the project will entail optimizing large-scale production of protein transduced long-term hematopoietic stem cells from human cells that are resistant to HIV infection for eventual testing in larger animals and human clinical trials. PUBLIC HEALTH RELEVANCE: This project seeks to optimize a novel method to generate large amounts of human blood stem cells for clinical use in the prevention and treatment of HIV infection. Once this method has been validated for large-scale production, the long-term goal for benefiting public health will be to use this system to generate HIV resistant cells in infected individuals or in populations that are at-risk for acquiring the HIV virus.